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1.
This report describes a case of cutaneous pythiosis in a 6‐year‐old female mixed breed dog, from the central west region of São Paulo State, Brazil. The cytological and histopathological analyses showed an intense inflammatory infiltrate with presence of numerous hyphal elements, suggesting infection due to Pythium insidiosum. The diagnosis was confirmed by nested‐PCR, which was carried out with specific primers derived from the ribosomal DNA region. The pathogen occurs in Brazil and veterinarians should be aware of the importance of correctly diagnosing this disease and differentiating it from other fungal diseases.  相似文献   

2.
Abstract: A 2‐year‐old, castrated male, mixed‐breed dog was presented to the University of Florida Veterinary Medical Center with swelling, edema, ulceration, and draining tracts in the region surrounding the left hock. The dog had mild monocytosis and moderate hyperglobulinemia. Fine‐needle aspirate specimens of the left popliteal lymph node revealed pyogranulomatous lymphadenitis with hyphal organisms. The diameters of the hyphae were variable, ranging from 11 to 22 μm. The organism was considered as most consistent with Lagenidium caninum; although Pythium insidiosum or Lagenidium karlingii were not conclusively excluded, hyphal diameter in these organisms is typically smaller (6.6–8.8 and 2.5–11 μm, respectively). A positive Western blot confirmed the presence of serum antibodies reactive against Lagenidium sp. and the absence of antibodies to P. insidoisum, Basidiobolus, and Conidiobolus antibodies. Careful assessment of hyphal diameter in cytologic specimens may be useful in differentiating L. caninum from P. insidiosum or L. karlingii.  相似文献   

3.
Genotyping of Mycobacterium avium subspecies paratuberculosis (MAP) is important for precise classification of bacterium and for understanding the molecular epidemiology. The present study reports detection and typing of the MAP from milk. On the basis of clinical signs of diarrhea and/or weakness, the dairy animals suspected for Johne’s disease were screened by Ziehl–Neelsen staining of fecal samples. The milk samples from 13 selected animals were processed for DNA extraction and direct IS900 polymerase chain reaction (PCR). MAP identified by IS900 PCR was genotyped using IS1311 PCR-restriction endonuclease analysis (REA). IS900 milk PCR revealed 30.8% animals positive for MAP, including 40% of the moderate and 50% of the heavy fecal shedders. All infected animals showed Bison type MAP in IS1311 PCR-REA. IS900 PCR can be used for screening of milk for MAP; however, the method needs to be evaluated for subclinical cases. IS1311 PCR-REA results indicated the predominance of Bison type MAP in the dairy animals of this region.  相似文献   

4.
Abstract The purpose of this study was to evaluate the application of previously described Pythium insidiosum‐ and Lagenidium‐specific nested PCR assays to the detection of oomycete DNA in animal tissues. DNA was extracted from 15 frozen and 10 ethanol‐fixed tissues obtained from six animals with pythiosis, five animals with lagenidiosis, one animal with nonoomycotic skin disease and two animals without skin disease. First‐round PCR, which utilized universal fungal primers ITS1 and ITS2P, amplified a single product of the expected size for each of the P. insidiosum‐ and Lagenidium‐infected tissues, but not for tissues obtained from animals without fungal disease. Second‐round PCR using the P. insidiosum‐specific primers PI1 and PI2 produced a single 105‐bp product for the P. insidiosum‐infected tissues, but not for any of the other tissues. Second‐round PCR using the Lagenidium‐specific primers LAG1 and LAG2 produced a single 76‐bp product for the Lagenidium‐infected tissues, but not for any of the other tissues.  相似文献   

5.
A mixed‐breed dog presented with tenesmus, hematochezia, and abdominal distension of 2 weeks duration. Radiography showed a large round mass with a “soap‐bubble” appearance and shell‐like mineralization in the caudal abdomen. Computed tomography revealed a lamellate mineralized mass 8 cm in diameter and containing air in the descending colon and prostatic abscess. Heterogeneously contrast‐enhanced, irregularly thickened colonic wall with intramural and peritoneal free gas indicated stercoral colonic perforation. Surgical intervention revealed a tumor‐like giant fecaloma in the descending colon adjoining the prostate with extensive wall rupture and fecal peritonitis. Hypothetically, prostatic inflammation may affect colonic motility with resultant fecaloma formation.  相似文献   

6.
Routine necropsies of 27 asymptomatic juvenile chinchillas revealed a high prevalence of gastric ulcers with microscopic lymphoplasmacytic gastroenteritis and typhlocolitis. Polymerase chain reaction (PCR) analysis using Campylobacter genus‐specific partial 16S rRNA primers revealed the presence of Campylobacter spp. DNA in the faeces of 12 of 27 animals (44.4%). Species‐specific partial 16S rRNA PCR and sequencing confirmed that these animals were colonized with Campylobacter lanienae, a gram‐negative, microaerophilic bacterium that was first identified on routine faecal screening of slaughterhouse employees and subsequently isolated from faeces of livestock. Campylobacter lanienae was isolated from the faeces of six PCR‐positive animals and identified with species‐specific PCR and full 16S rRNA sequencing. Phylogenetic analysis showed that these isolates clustered with C. lanienae strain NCTC 13004. PCR analysis of DNA extracted from gastrointestinal tissues revealed the presence of C. lanienae DNA in the caecum and colon of these chinchillas. Gastrointestinal lesions were scored and compared between C. lanienae‐positive and C. lanienae‐negative animals. There was no correlation between colonization status and lesion severity in the stomach, liver, duodenum, or colon. Possible routes of C. lanienae infection in chinchillas could include waterborne transmission and faecal–oral transmission from wild mice and rats or livestock. Based on these findings, the authors conclude that C. lanienae colonizes the lower bowel of chinchillas in the absence of clinical disease. This is the first report of C. lanienae in any rodent species. Campylobacter lanienae isolates from different mammalian species demonstrate heterogeneity by 16S rRNA sequence comparison. Analysis using rpoB suggests that isolates and clones currently identified as C. lanienae may represent multiple species or subspecies.  相似文献   

7.
Fecal DNA analysis is a useful tool for the investigation of endangered species. Tamaraw (Bubalus mindorensis) is endemic to the Philippine island of Mindoro but knowledge of its genetic and ecological information is limited. In this study, we developed a species identification method for tamaraw by fecal DNA analysis. Eighteen feces presumed to be from tamaraw were collected in Mount Iglit‐Baco National Park and species‐known feces from domestic buffaloes and cattle were obtained from a farm. Additionally, one species‐unknown fecal sample was obtained in Mount Aruyan Preserve, where the sighting of tamaraw has not been reported in recent years. Based on DNA sequence data previously reported, the genus Bubalus‐ and tamaraw‐specific primers for PCR of cytochrome b gene were newly designed. The Bubalus‐specific primer yielded a 976 bp fragment of cytochrome b for all fecal samples from tamaraw and domestic buffaloes, but not for cattle, whereas the tamaraw‐specific primer yielded a 582 bp fragment for all tamaraw fecal samples and for one of the four domestic buffalo samples. PCR‐RFLP (restriction fragment length polymorphism) analysis of the 976 bp PCR fragment with AvrII or BsaXI provided distinct differences between tamaraw and domestic buffalo. PCR‐RFLP analysis also showed that the species‐unknown sample obtained in Mount Aruyan Preserve, originates from tamaraw.  相似文献   

8.
The objective of this study is the detection of Mycobacterium avium subsp. paratuberculosis (MAP) by serum enzyme-linked immunosorbent assay (ELISA), fecal polymerase chain reaction (PCR), and fecal culture in Colombian dairy herds. Serum and fecal samples from asymptomatic cows (n = 307) of 14 dairy herds were tested for MAP by an unabsorbed ELISA test (ELISA-A). Serum and fecal samples from positive ELISA-A animals (n = 31) were further tested by an absorbed ELISA test (ELISA-B) and PCR. Fecal samples from animals of herds positive by ELISA-A and PCR (n = 105) were inoculated onto three different culture media. ELISA-A produced positive results in 10% of the serum samples and 71% of the herds. ELISA-B and PCR results were positive in two and six serum and fecal samples from positive ELISA-A animals, respectively. Fecal samples were negative for MAP on all culture media. The results of this study confirmed the presence of MAP in local dairy herds and the difficulties of MAP detection in asymptomatic animals by ELISA, PCR, and fecal culture.  相似文献   

9.
Biofilm production by Staphylococcus aureus, an important virulence factor was investigated employing phenotypic and genotypic methods. A total of 102 S. aureus isolates from bovine subclinical mastitis cases were included in the study. Maximum number of biofilm producing strains were detected by Congo red agar (CRA) method (48.03%) followed by tube method (36.27%). Tissue culture plate method (TCP) without and with destaining identified 19.60 and 29.41% of S. aureus as biofilm producers, respectively. A polymerase chain reaction for detection of intercellular adhesion genes, icaA and icaD, responsible for biofilm formation was standardized. Of the 102 S. aureus isolates investigated, 36 (35.29%) strains revealed presence of both the genes. Considering polymerase chain reaction as a standard test, CRA and TCP without destaining were the most sensitive and specific, respectively. PCR technique standardized for detection of the icaA and icaD genes is reliable for identifying biofilm producing potential of S. aureus which may help in rapid detection of biofilm-producer Staphylococci. This would allow the early application of control measures.  相似文献   

10.
Background: Measurement of proteolytic activity in feces is a traditional method for the diagnosis of exocrine pancreatic insufficiency (EPI). A drawback of this method is the occurrence of falsely low results that may lead to a false‐positive diagnosis of EPI. We hypothesized that intestinal loss of serum proteinase inhibitors in protein‐losing enteropathy (PLE) may inhibit fecal proteolytic activity and be a potential source of false low results. Objective: The objective of this study was to determine the effect of PLE on fecal proteolytic activity in dogs. Methods: Fecal proteolytic activity was measured using a radial diffusion casein digestion assay in 12 samples from 4 clinically healthy control dogs and 30 samples from 16 dogs with PLE. Gastrointestinal protein loss was assessed using an ELISA to determine fecal canine α1‐proteinase inhibitor concentration. The relationship between the concentration of canine α1‐proteinase inhibitor in the feces and the diameter cleared in the casein digestion assay was determined. The mean clearing diameter was compared between control dogs and dogs with PLE. Results: A significant negative correlation was observed between fecal canine α1‐proteinase inhibitor concentration and casein clearing diameter (P < .001, Pearson r=—.6317, r 2 =.3999). Mean clearing diameter was significantly lower in dogs with PLE than in control dogs (12.63 vs 16.83 mm, P < .001, two‐tailed Student's t‐test). Conclusion: Increased fecal loss of α1‐proteinase inhibitor in dogs with PLE is associated with a significant decrease in fecal proteolytic activity and may result in a false positive diagnosis of EPI.  相似文献   

11.
Faecal samples obtained from 190 healthy mithuns were examined for the presence of Escherichia coli. Total one‐hundred and five E. coli isolates were obtained from these samples, which belonged to 55 different serogroups. These isolates were subjected to multiplex polymerase chain reaction (m‐PCR) for detection of stx1, stx2, eaeA and hlyA genes. Twenty‐three (21.90%) E. coli isolates belonging to 14 serogroups revealed the presence of at least one virulence gene when examined by m‐PCR. Nineteen percent and 2.85% of the mithuns were found to carry Shiga toxin‐producing E. coli (STEC) and enteropathogenic E. coli, respectively. stx1 and stx2 genes were found to be prevalent in 7 (6.67%) and 18 (17.14%) of the isolates respectively, whereas eaeA and hlyA genes were found to be carried by three (2.85% each) isolates. Interestingly, none of the STEC isolates belonged to serogroup O157.  相似文献   

12.
A 20-month-old castrated male Labrador Retriever with a 3-month history of anorexia, weight loss, and vomiting was evaluated. Plasma biochemical abnormalities included marked hyperglobulinemia and hypercalcemia. Serum levels of parathyroid hormone, parathyroid hormone-related protein, 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D were either low or within reference intervals. Gastric wall thickening and abdominal lymphadenomegaly were observed with abdominal ultrasonography. Cytologic evaluation of a sample obtained via fine-needle aspiration of the gastric wall revealed pyogranulomatous inflammation and numerous poorly stained hyphae. Partial gastrectomy was performed, and a diagnosis of gastric pythiosis was made by immunohistochemical staining of infected gastric tissue, as well as by immunoblot serology. This case demonstrates that diagnostic samples for cytologic evaluation can be obtained by fine-needle aspiration of Pythium insidiosum-infected tissues and that a presumptive diagnosis can be made by examination of a Romanowsky-stained smear. Furthermore, pythiosis should be considered as a differential diagnosis for hypercalcemia, especially in young dogs with inflammatory lesions that have a granulomatous component. The mechanism for the hypercalcemia in this dog was not determined; however, calcium concentrations normalized after surgical resection of the gastric lesion.  相似文献   

13.
Tritrichomonas foetus is a venereal pathogen of naturally bred cattle. In domestic cats, T. foetus colonizes the colon, resulting in chronic, large-bowel diarrhea. The infection is prevalent among young, densely housed cats, and there is no effective treatment. To the authors' knowledge, the characteristic microscopic lesions of T. foetus infection in naturally infected cats have not been described. The aim of the study reported here was to characterize the histologic changes in the colon of seven cats with T. foetus infection and chronic diarrhea. All cats were 1 year old or younger (mean, 6.7 +/- 1.7 months), and a diagnosis of T. foetus infection was made on the basis of direct fecal smear examination (five cats), fecal culture in InPouch TF medium (four cats), single-tube nested polymerase chain reaction (PCR) analysis of DNA extracted from feces (two cats), or observation of trichomonads in sections of colon followed by PCR confirmation on DNA extracted from paraffin-embedded tissue (two cats). The presence of colonic trichomonads was the most diagnostic histologic feature. Organisms were identified in all cats, but in only 24 of 43 (56%) sections of colon. Trichomonads were generally present in close proximity to the mucosal surface and less frequently in the lumen of colonic crypts. The presence of colonic trichomonads was consistently associated with mild-to-moderate lymphoplasmacytic and neutrophilic colitis, crypt epithelial cell hypertrophy, hyperplasia and increased mitotic activity, loss of goblet cells, crypt microabscesses, and attenuation of the superficial colonic mucosa. In two of the cats, histologic lesions were more severe and were associated with invasion of trichomonads into the lamina propria and/or deeper layers of the colon.  相似文献   

14.
Gastrointestinal (GI) pythiosis is a severe and often fatal disease in dogs that traditionally has been poorly responsive to medical treatment. Although aggressive surgical resection with wide margins is the most consistently effective treatment, lesion location and extent often preclude complete resection. Recently, it has been suggested that the addition of anti‐inflammatory doses of corticosteroids may improve outcome in dogs with nonresectable GI pythiosis. This report describes 3 dogs with colonic pythiosis in which complete resolution of clinical signs, regression of colonic masses, and progressive decreases in serological titers were observed after treatment with itraconazole, terbinafine, and corticosteroids. This treatment protocol represents a promising treatment for dogs with GI pythiosis in which surgical intervention is not feasible.  相似文献   

15.
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16.
Background: Salmon poisoning disease (SPD) is a trematode‐borne disease of dogs caused by Neorickettsia helminthoeca. Objectives: To determine risk factors and spatial epidemiology of SPD in dogs from northern California; to describe the clinicopathologic, microbiologic, and imaging findings of SPD in these dogs; and to evaluate treatments and outcomes for SPD. Animals: Twenty‐nine dogs with SPD based on the finding of trematode ova in the feces, or organisms consistent with N. helminthoeca in specimens submitted for microscopic examination. Methods: Information regarding signalment, fish exposure, clinical signs, diagnostic evaluation, treatments, and outcomes was obtained for each dog. Archived lymph node aspirates and histopathology specimens were subjected to polymerase chain reaction (PCR) testing for Neorickettsia spp. Results: Labrador Retrievers and intact male dogs were overrepresented. Exposure locations were often distant from the dogs' residence. Some dogs had neurologic signs, including twitching and seizures. Dogs lacking peripheral lymphadenomegaly had abdominal lymphadenomegaly on ultrasound examination. A combination of centrifugation fecal flotation and sedimentation had greatest sensitivity for finding fluke ova. N. helminthoeca DNA was amplified by PCR from 4/10 dogs. Penicillins, cephalosporins, and chloramphenicol did not appear to be effective treatments. Mortality rate was 4/29 (14%). Conclusions and Clinical Importance: SPD should be suspected in dogs with inappetence, gastrointestinal, or neurologic signs, with or without fever or peripheral lymphadenomegaly in the appropriate geographical setting. Diagnosis is facilitated by a combination of fecal sedimentation and centrifugal flotation, abdominal ultrasonography, and PCR‐based assays on lymphoid tissue. The treatment of choice is tetracycline antimicrobials.  相似文献   

17.
Here, we describe the establishment of mutant‐specific polymerase chain reaction (PCR) for detection of a c‐KIT c.1430G>T mutation in feline mast cell tumours. Several mutations in feline c‐KIT have been identified, with the c.1430G>T mutation accounting for a significant portion of feline mast cell tumour mutations. The c.1430G>T mutation in c‐KIT exon 9 was detected in 15.7% (11 of 70) of samples by mutant‐specific PCR but in only 7.1% (5 of 70) by PCR–restriction fragment length polymorphism (RFLP) in the genomic DNA isolated from 70 formalin‐fixed paraffin‐embedded sections or cells collected by fine needle aspiration. Mutant‐specific PCR showed remarkably higher detection rate than did PCR–RFLP. DNA sequence analysis did not always yield identical results to those of mutant‐specific PCR, suggesting heterogeneity of tumour cells. Mutant‐specific PCR is a valid and efficient screening tool for detection of the c‐KIT c.1430G>T point mutation in feline mast cell tumours compared with PCR–RFLP and sequencing analysis.  相似文献   

18.
Fecal specimens were obtained from a total of 70 dairy calves less than two months old on 11 municipalities in Buenos Aires, Argentina. After removal of fecal debris by sieving and sucrose flotation, specimens were subjected to PCR to detect the presence of Enterocytozoon bieneusi. PCR revealed a 14.3% of prevalence for E. bieneusi with 10 positive calves from 7 municipalities. Gene sequence analysis conducted in all samples positives by PCR revealed the presence of six genotypes; four previously reported in cattle as well as humans (D, I, J, and BEB4), one never reported in cattle before but previously reported in humans (EbpC), and one novel genotype (BEB10). These results constitute the first molecular characterization of E. bieneusi in Argentina, and suggest a potential risk of zoonotic transmission in this area.  相似文献   

19.
One hundred and fifty‐eight staphylococcal strains isolated from wild rodents and insectivores were analysed for plasmid‐borne resistance to tetracycline (Tc). Only 10 isolates, six Staphylococcus saprophyticus isolates and single isolates of S. xylosus, S. equorum, S. warneri and S. cohnii subsp. cohnii carried a Tc resistance plasmid of approximately 4.4 kb as confirmed by protoplast transformation. All 10 plasmids harboured a Tc resistance gene of hybridization class K [tet(K)] as confirmed by polymerase chain reaction (PCR). The plasmid was assigned to the pT181 family as it revealed a high degree of restriction map homology to pT181 and other members of this family. Macrorestriction analysis with the enzyme SmaI showed that three of the six isolates identified as S. saprophyticus shared the same pulsed‐field gel electrophoresis (PFGE) pattern.  相似文献   

20.
In the present article the distribution and abundance of ammonia‐assimilating microbes among the natural habitants in a lagoon were investigated. In the medium containing lagoon‐extract, about 20% of total 82 isolates showed ammonia‐assimilating ability. By sequencing of 16S ribosomal DNA (rDNA), the highest ammonia‐assimilating isolates at 10°C and 37°C were identified as Janthinobacterium lividum and Bacillus sp., respectively. The structure of the microbial community was analyzed by polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE). Almost of the dominant species that were detected by PCR‐DGGE did not coincide with isolates, which showed the high ammonia‐assimilating ability, but one specie by PCR‐DGGE coincided with ammonia‐assimilating isolate. These results suggested that ammonia‐assimilating microbes existed as non‐dominant species in the microbial community in a lagoon.  相似文献   

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